Fig. 3: Ag⁺ CD4⁺ and CD8⁺ T cell functionality.

A Percentage of Ag⁺ CD4⁺ T cells producing different cytokines after in vitro stimulation with SARS-CoV-2 peptides. Representative dot plots showing the percentages of CD4⁺ Ag⁺ cells producing IL-2, IL-17, CD107a and granzyme B (GRZB). Plots show mean (center bar) ± SD. Kruskal–Wallis (one-sided) test with Benjamini-Hochberg correction for multiple comparisons. B Polyfunctional profile of Ag⁺ CD4⁺ T cells. (Upper) Pie charts representing the proportion of Ag⁺ CD4⁺ T cells producing different combinations of CD107a, IL2, IL17, IFNγ, and TNF. Each color refers to specific polyfunctional CD4 T subpopulation as reported in the ‘polyfunctionality legend’. The far-left heatmap illustrates the statistical variances among the 9 distinct pie charts; The central bar represents the mean ± SD. Kruskal–Wallis test (one-sided) with Benjamini-Hochberg correction for multiple comparisons. (far-right) Dot plot reporting the percentages of Ag⁺ CD4⁺ producing different combination of cytokines. Kruskal-Wallis (one-sided) test with Benjamini–Hochberg correction for multiple comparisons. C Percentage of Ag⁺ CD8⁺ T cells producing different cytokines after in vitro stimulation with SARS-CoV-2 peptides. Representative dot plots of Ag⁺ CD8⁺ cells producing CD107a and GRZB. Dot plot representing the percentage of Ag⁺ CD8⁺ T cells producing GRZB is shown, mean (center bar) ± SD. Kruskal–Wallis test (one-sided) with Benjamini–Hochberg correction for multiple comparisons. D Polyfunctional profile of Ag⁺ CD8⁺ T cells. (Upper) Pie charts representing the percentage of Ag⁺ CD8⁺ T cells producing different combinations of CD107a, IL2, IL17, IFNγ, and TNF. Each color refers to specific polyfunctional CD8 T subpopulation as reported in the ‘polyfunctionality legend’. The far-left heatmap illustrates the statistical variances among the 9 distinct pie charts; Kruskal-Wallis test (one-sided) with Benjamini–Hochberg correction for multiple comparisons (Right) Dot plot reporting the percentage of Ag⁺ CD8⁺ CD107a⁻IFNγ⁻IL2⁻IL17⁻TNF population. The central bar represents the mean ± SEM. Kruskal–Wallis test with Benjamini–Hochberg correction for multiple comparisons was used to test the differences among the nine groups. In A–C plots: HD healthy donors (N = 13); Cladribine (N = 6); DMF Dimethyl Fumarate (N = 14); DMF Lymphopenic: Dimethyl Fumarate Lymphopenic (N = 9); Fingolimod (N = 12); IFN Interferon 1β (N = 13); Natalizumab (N = 15); Teriflunomide (N = 8); rituximab/ocrelizumab (N = 11).